Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis.
The dimeric nature of triosephosphate isomerases (TIMs) is maintained by an extensive surface area interface of more than 1600 Å2. TIMs from Trichomonas vaginalis (TvTIM) are held in their dimeric state by two mechanisms: a ball and socket interaction of residue 45 of one subunit that fits into the...
| Main Authors: | , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2015-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC4664265?pdf=render |
| _version_ | 1818566697694527488 |
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| author | Samuel Lara-Gonzalez Priscilla Estrella Carmen Portillo María E Cruces Pedro Jimenez-Sandoval Juliana Fattori Ana C Migliorini-Figueira Marisol Lopez-Hidalgo Corina Diaz-Quezada Margarita Lopez-Castillo Carlos H Trasviña-Arenas Eugenia Sanchez-Sandoval Armando Gómez-Puyou Jaime Ortega-Lopez Rossana Arroyo Claudia G Benítez-Cardoza Luis G Brieba |
| author_facet | Samuel Lara-Gonzalez Priscilla Estrella Carmen Portillo María E Cruces Pedro Jimenez-Sandoval Juliana Fattori Ana C Migliorini-Figueira Marisol Lopez-Hidalgo Corina Diaz-Quezada Margarita Lopez-Castillo Carlos H Trasviña-Arenas Eugenia Sanchez-Sandoval Armando Gómez-Puyou Jaime Ortega-Lopez Rossana Arroyo Claudia G Benítez-Cardoza Luis G Brieba |
| author_sort | Samuel Lara-Gonzalez |
| collection | DOAJ |
| description | The dimeric nature of triosephosphate isomerases (TIMs) is maintained by an extensive surface area interface of more than 1600 Å2. TIMs from Trichomonas vaginalis (TvTIM) are held in their dimeric state by two mechanisms: a ball and socket interaction of residue 45 of one subunit that fits into the hydrophobic pocket of the complementary subunit and by swapping of loop 3 between subunits. TvTIMs differ from other TIMs in their unfolding energetics. In TvTIMs the energy necessary to unfold a monomer is greater than the energy necessary to dissociate the dimer. Herein we found that the character of residue I45 controls the dimer-monomer equilibrium in TvTIMs. Unfolding experiments employing monomeric and dimeric mutants led us to conclude that dimeric TvTIMs unfold following a four state model denaturation process whereas monomeric TvTIMs follow a three state model. In contrast to other monomeric TIMs, monomeric variants of TvTIM1 are stable and unexpectedly one of them (I45A) is only 29-fold less active than wild-type TvTIM1. The high enzymatic activity of monomeric TvTIMs contrast with the marginal catalytic activity of diverse monomeric TIMs variants. The stability of the monomeric variants of TvTIM1 and the use of cross-linking and analytical ultracentrifugation experiments permit us to understand the differences between the catalytic activities of TvTIMs and other marginally active monomeric TIMs. As TvTIMs do not unfold upon dimer dissociation, herein we found that the high enzymatic activity of monomeric TvTIM variants is explained by the formation of catalytic dimeric competent species assisted by substrate binding. |
| first_indexed | 2024-12-14T01:56:55Z |
| format | Article |
| id | doaj.art-427e6c8416424f5c82dfbcc25abb4acf |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T01:56:55Z |
| publishDate | 2015-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-427e6c8416424f5c82dfbcc25abb4acf2022-12-21T23:21:10ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011011e014174710.1371/journal.pone.0141747Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis.Samuel Lara-GonzalezPriscilla EstrellaCarmen PortilloMaría E CrucesPedro Jimenez-SandovalJuliana FattoriAna C Migliorini-FigueiraMarisol Lopez-HidalgoCorina Diaz-QuezadaMargarita Lopez-CastilloCarlos H Trasviña-ArenasEugenia Sanchez-SandovalArmando Gómez-PuyouJaime Ortega-LopezRossana ArroyoClaudia G Benítez-CardozaLuis G BriebaThe dimeric nature of triosephosphate isomerases (TIMs) is maintained by an extensive surface area interface of more than 1600 Å2. TIMs from Trichomonas vaginalis (TvTIM) are held in their dimeric state by two mechanisms: a ball and socket interaction of residue 45 of one subunit that fits into the hydrophobic pocket of the complementary subunit and by swapping of loop 3 between subunits. TvTIMs differ from other TIMs in their unfolding energetics. In TvTIMs the energy necessary to unfold a monomer is greater than the energy necessary to dissociate the dimer. Herein we found that the character of residue I45 controls the dimer-monomer equilibrium in TvTIMs. Unfolding experiments employing monomeric and dimeric mutants led us to conclude that dimeric TvTIMs unfold following a four state model denaturation process whereas monomeric TvTIMs follow a three state model. In contrast to other monomeric TIMs, monomeric variants of TvTIM1 are stable and unexpectedly one of them (I45A) is only 29-fold less active than wild-type TvTIM1. The high enzymatic activity of monomeric TvTIMs contrast with the marginal catalytic activity of diverse monomeric TIMs variants. The stability of the monomeric variants of TvTIM1 and the use of cross-linking and analytical ultracentrifugation experiments permit us to understand the differences between the catalytic activities of TvTIMs and other marginally active monomeric TIMs. As TvTIMs do not unfold upon dimer dissociation, herein we found that the high enzymatic activity of monomeric TvTIM variants is explained by the formation of catalytic dimeric competent species assisted by substrate binding.http://europepmc.org/articles/PMC4664265?pdf=render |
| spellingShingle | Samuel Lara-Gonzalez Priscilla Estrella Carmen Portillo María E Cruces Pedro Jimenez-Sandoval Juliana Fattori Ana C Migliorini-Figueira Marisol Lopez-Hidalgo Corina Diaz-Quezada Margarita Lopez-Castillo Carlos H Trasviña-Arenas Eugenia Sanchez-Sandoval Armando Gómez-Puyou Jaime Ortega-Lopez Rossana Arroyo Claudia G Benítez-Cardoza Luis G Brieba Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis. PLoS ONE |
| title | Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis. |
| title_full | Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis. |
| title_fullStr | Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis. |
| title_full_unstemmed | Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis. |
| title_short | Substrate-Induced Dimerization of Engineered Monomeric Variants of Triosephosphate Isomerase from Trichomonas vaginalis. |
| title_sort | substrate induced dimerization of engineered monomeric variants of triosephosphate isomerase from trichomonas vaginalis |
| url | http://europepmc.org/articles/PMC4664265?pdf=render |
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